Glyosylation markers for cancer and chronic inflammation

Abstract

The present invention provides novel biomarkers for use in the diagnosis and prognosis of cancerous and malignant conditions and further of diseases which are mediated by a proinflammatory immune response. The biomarkers are glycoproteins, the levels of expression of which have been correlated by the inventors to correspond to particular disease conditions. The invention further extends to methods for use in monitoring the response to therapy of a treatment for use in the treatment of a cancerous condition or proinflammatory disease.

1 . A method for the diagnosis of a cancerous and/or malignant condition, the prognosis of a cancerous and/or malignant condition, and/or the monitoring of a response to treatment of a cancerous and/or malignant condition in a subject, the method comprising the steps of:—
 providing a test sample from the subject, 
 determining a level in the test sample of two or more glycosylation markers for the cancerous and/or malignant condition, 
 providing a diagnosis, prognosis or determination of the response based on the level of the two or more glycosylation markers. 
 
     
     
         2 . A method as claimed in  claim 1 , wherein the level of 2, 3, 4, 5, 6, 7, 8, 9 or 10 glycosylation markers for the cancerous and/or malignant condition are determined. 
     
     
         3 . A method for the diagnosis of a cancerous and/or malignant condition, the prognosis of a cancerous and/or malignant condition and/or the monitoring of a response to treatment of a cancerous and/or malignant condition in a subject, the method comprising the steps of:
 providing a test sample from the subject,   determining the level in the test sample of one or more glycosylation marker(s) of the cancerous and/or malignant condition and one or more non-glycosylation marker(s) of the cancerous and/or malignant condition, and   providing a diagnosis, prognosis or determination of the response based on the level of the one or more glycosylation markers and the one or more non-glycosylation markers.   
     
     
         4 . A method as claimed in  claim 3 , wherein the level of 2, 3, 4, 5, 6, 7, 8, 9 or 10 glycosylation markers for the cancerous and/or malignant condition are determined. 
     
     
         5 . A method as claimed in  claim 3 , wherein the level of 2, 3, 4, 5, 6, 7, 8, 9 or 10 non-glycosylation markers for the cancerous and/or malignant condition are determined. 
     
     
         6 . A method as claimed in  claim 3 , wherein the non-glycosylation markers are selected from the group consisting of inflammatory markers, cytokines, chemokines, genetic markers, Catecholamines, Immunoglobulins, markers for angiogenesis, or the like, or any combination thereof. 
     
     
         7 . A method as claimed in  claim 3 , wherein the non-glycosylation markers are selected from the group consisting of Alphafetoprotein, NMP22, Carcinoembryonic antigen (CEA), HER-2, CA 15-3, CA 27-29, CA 125, CA 19-9, and C-reactive protein (CRP), IL-4, IL-10, IL-1α and IL-1β, MCP-1, or the like, or any combination thereof. 
     
     
         8 . A method for the diagnosis of a cancerous and/or malignant condition, the prognosis of a cancerous and/or malignant condition and/or the monitoring of a response to treatment of a cancerous and/or malignant condition in a subject, the method comprising the steps of:
 providing a test sample from the subject,   determining the level of at least one marker selected from the group comprising glycans with GU values greater than 10.65, SLe x  structures, A2FG1 derived from digestion of SLe x , A3FG1 derived from digestion of SLe x , A4FG1 derived from digestion of SLe x , sialylated tri-antennary glycans, sialylated tetra-antennary glycans, glycans containing α1,3 fucose, α1,3 monofucosylated tri-antennary glycans, α1,3 difucosylated tri-antennary glycans, α1,3 monofucosylated tetra-antennary glycans, α1,3 difucosylated tetra-antennary glycans, tetra-antennary glycans with lactosamine extensions, ratio of α2,3 sialylated glycans to α2,6 sialylated glycans, agalactosylated fucosylated biantennary glycans, core fucosylated agalactosylated biantennary glycans, core fucosylated monosialylated glycans on transferrin, SLe x  on glycans on haptoglobin β-chain, A3FG1 derived from digestion of SLe x  on glycans on haptoglobin β-chain, A4FG1 derived from digestion of SLe x  on glycans on haptoglobin β-chain, SLe x  on glycans on α1-acid glycoprotein, A3FG1 derived from digestion of SLe x  on glycans on α1-acid glycoprotein, SLe x  on glycans on α1-antichymotrypsin, A3FG1 derived from digestion of SLe x  on glycans on α1-antichymotrypsin, tetra-antennary tetragalactosylated glycans on α1-antitrypsin, core fucosylated agalactosylated biantennary glycans on IgG, agalactosylated glycans on IgG, sialylation on glycans on IgG, galactosylation on glycans on IgG, FA2G2S1 on glycans on transferrin, FA2BG2S1 on glycans on transferrin and A4G4 glycans on α1-antitrypsin, or the like, or any combination thereof, and   providing a diagnosis, prognosis or determination of the response based on the determined level of the at least one marker.   
     
     
         9 . A method as claimed in  claim 1 , wherein the glycosylation markers are selected from the group consisting of changes in glycan branching; changes in levels of oligomannose, of hybrid and complex type N-glycans, of O-glycans, or of components thereof; changes in ratios of levels between glycans, GU values; or the like; or any combination thereof. 
     
     
         10 . A method as claimed in  claim 1 ,
 wherein the glycosylation markers are selected from the group consisting of: glycans with GU values greater than 10.65, SLe x  structures, A2FG1 derived from digestion of SLe x , A3FG1 derived from digestion of SLe x , A4FG1 derived from digestion of SLe x , sialylated tri-antennary, sialylated tetra-antennary glycans, glycans containing α1,3 fucose, α1,3 monofucosylated tri-antennary glycans, α1,3 difucosylated tri-antennary glycans, α1,3 monofucosylated tetra-antennary glycans, α1,3 difucosylated tetra-antennary glycans, tetra-antennary glycans with lactosamine extensions, ratio of α2,3 sialylated glycans to α2,6 sialylated glycans, agalactosylated fucosylated biantennary glycans, core fucosylated agalactosylated biantennary glycans, core fucosylated monosialylated glycans on transferrin, SLe x  on glycans on haptoglobin β-chain, A3FG1 derived from digestion of SLe x  on glycans on haptoglobin β-chain, A4FG1 derived from digestion of SLe x  on glycans on haptoglobin β-chain, SLe x  on glycans on α1-acid glycoprotein, A3FG1 derived from digestion of SLe x  on glycans on α1-acid glycoprotein, SLe x  on glycans on α1-antichymotrypsin, A3FG1 derived from digestion of SLe x  on glycans on α1-antichymotrypsin, tetra-antennary tetragalactosylated glycans on α1-antitrypsin, core fucosylated agalactosylated biantennary glycans on IgG, agalactosylated glycans on IgG, sialylation on glycans on IgG, galactosylation on glycans on IgG, FA2G2S1 on glycans on transferrin, FA2BG2S1 on glycans on transferrin and A4G4 on glycans on α1-antitrypsin, or the like, or any combination thereof.   
     
     
         11 . The method as claimed in  claim 1 , wherein the method involves the analysis of all members of one of the following groups of glycosylation markers S3 and S4, fucose, GU of 10.65 and tri and tetra antennary glycans; A3FG1 and FA2; SLe x  and fucosylated agalactosylated biantennary glycans; or the like; or combinations thereof. 
     
     
         12 . A method as claimed in  claim 3 , wherein the method involves the analysis of all members of one of the following groups of markers CRP and any one, two, three, four, five, six, or more of any glycosylation marker(s); CRP and any of one, two or three of the glycosylation markers S3 and S4, fucose, GU of 10.65, tri and tetra-antennary glycans; S3 and S4, fucose, GU of 10.65, tri and tetra-antennary glycans, and CRP; fucosylated agalactosylated biantennary glycans and CRP; one or more pro-inflammatory cytokines and one or more glycosylation marker; one or more anti-inflammatory cytokine and one or more glycosylation marker; one or more chemokine and one or more glycosylation marker; or the like; or combinations thereof. 
     
     
         13 . The method as claimed in  claim 1 , wherein the subject is a human. 
     
     
         14 . The method as claimed in  claim 1 , wherein the test sample comprises a body fluid or body tissue. 
     
     
         15 . The method as claimed in  claim 14 , wherein the body fluid comprises serum, plasma or urine. 
     
     
         16 . The method as claimed in  claim 1 , comprising comparing the level of the one or more markers in the test sample with a level of the one or more markers in a control sample to determine the diagnosis, prognosis, and/or response. 
     
     
         17 . The method as claimed in  claim 16 , wherein an increase in the level of the one or more markers in the test sample as compared to the control sample indicates the presence of cancer. 
     
     
         18 . The method as claimed in  claim 1 , wherein determining the level in the test sample of the two or more markers comprises determining the level of the two or more markers on one or more acute phase proteins in the test sample. 
     
     
         19 . The method of  claim 18 , wherein the one or more acute phase proteins are selected from the group consisting of: serum amyloid A, haptoglobin, α1-acid glycoprotein, α1-antitrypsin, α1-antichymotrypsin, fibrinogen and transferrin. 
     
     
         20 . The method of  claim 18 , the method comprising isolating the one or more acute phase proteins from the test sample prior to determining the level of the two or more markers on the one or more acute phase proteins. 
     
     
         21 . The method as claimed in  claim 1 , wherein determining the level in the test sample of the two or more markers comprises releasing a pool of glycans from total glycoproteins in the test sample and determining the level of the two or more markers in the pool of glycans. 
     
     
         22 . The method as claimed in  claim 1 , wherein determining the level in the test sample of the two or more markers comprises performing chromatography on the sample or a derivative or component thereof. 
     
     
         23 . The method as claimed in  claim 1 , wherein determining the level in the test sample of the two or more markers comprises performing mass spectrometry, immuno-PCR, two-dimensional gel electrophoresis, ELISA, lectin ELISA, Western blot, immunoassay, lectin immunoassay, or one dimensional gel electrophoresis on the sample or a derivative or component thereof. 
     
     
         24 . A method for assessing the inflammatory state of a subject, the method comprising the steps of:—
 providing a test sample from a subject, 
 determining a level in the test sample of two or more glycosylation markers for chronic inflammation, 
 providing an assessment based on the level of the two or more glycosylation marker. 
 
     
     
         25 . A method for assessing the inflammatory state of a subject, the method comprising the steps of:
 providing a test sample from a subject,   determining the level in the test sample of one or more glycosylation marker(s) of chronic inflammation and one or more non-glycosylation marker(s) of chronic inflammation, and   providing an assessment based on the level of the one or more glycosylation markers and the one or more non-glycosylation markers.   
     
     
         26 . A method for assessing the inflammatory state of a subject, the method comprising the steps of:
 providing a test sample from a subject,   determining the level of at least one marker selected from the group comprising glycans with GU values greater than 10.65, SLe x  structures, A2FG1 derived from digestion of SLe x , A3FG1 derived from digestion of SLe x , A4FG1 derived from digestion of SLe x , sialylated tri-antennary glycans, sialylated tetra-antennary glycans, glycans containing α1,3 fucose, α1,3 monofucosylated tri-antennary glycans, α1,3 difucosylated tri-antennary glycans, α1,3 monofucosylated tetra-antennary glycans, α1,3 difucosylated tetra-antennary glycans, tetra-antennary glycans with lactosamine extensions, ratio of α2,3 sialylated glycans to α2,6 sialylated glycans, agalactosylated fucosylated biantennary glycans, core fucosylated agalactosylated biantennary glycans, core fucosylated monosialylated glycans on transferrin, SLe x  on glycans on haptoglobin β-chain, A3FG1 derived from digestion of SLe x  on glycans on haptoglobin β-chain, A4FG1 derived from digestion of SLe x  on glycans on haptoglobin β-chain, SLe x  on glycans on α1-acid glycoprotein, A3FG1 derived from digestion of SLe x  on glycans on α1-acid glycoprotein, SLe x  on glycans on α1-antichymotrypsin, A3FG1 derived from digestion of SLe x  on glycans on α1-antichymotrypsin, tetra-antennary tetragalactosylated glycans on α1-antitrypsin, core fucosylated agalactosylated biantennary glycans on IgG, agalactosylated glycans on IgG, sialylation on glycans on IgG, galactosylation on glycans on IgG, FA2G2S1 on glycans on transferrin, FA2BG2S1 on glycans on transferrin and A4G4 glycans on α1-antitrypsin, or the like, or any combination thereof, and   providing an assessment based on the determined level of the at least one marker.